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人大肠癌线粒体dna重组质粒的构建

, PP. 1016-1019

Keywords: 大肠癌,线粒体,dna,d-环区,突变,质粒

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Abstract:

目的了解大肠癌细胞株(sw480,lovo,ht29)线粒体dna的突变,克隆突变的大肠癌线粒体dna(mtdna)基因,构建pcdna3.i(+)-mtdna真核表达重组体,并导入nih3t3细胞,以探讨线粒体基因突变与肿瘤发生的关系。方法提取大肠癌细胞株(sw480,lovo,ht29)mtdna,扩增d-loop区,产物用dna自动测序法进行序列分析。利用dna重组技术将其定向插人真核表达质粒pcdna3.1(+),并用脂质体法导人nih3t3细胞。结果检测出大肠癌细胞株sw480、lovo、ht29细胞mtdnad-loop分别有10、9、8个突变位点。成功克隆1119bp的mtdnad-loop区至表达质粒pcdna3.1(+),并导入nih3t3细胞中。结论线粒体dnad-loop区是一个具有高度多态性和突变性的区域,在大肠癌细胞株中突变率较高。

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